Figure 4.

Stability of the mean lifetime during time-lapse measurement of the mCherry/EGFP and mTFP1/EYFP couple. Representative fast acquisitions of 15 s using the TriM-FLIM system are presented for the two couples. Identical regions of interest were chosen to calculate the average mean lifetime for each image of a time lapse of 20 acquisitions. The average mean lifetime for EGFP alone and mCherry-EGFP tandem was 2.45 and 2.23 ns, respectively (this decrease is stressed by the small arrow on the left-hand side of the graph). The average mean lifetime for the mTFP1alone and mTFP1-EYFP tandem was 2.71 and 2.08 ns, respectively (this decrease is stressed by the big arrow on the right-hand side of the graph).